Pulmonary microbial infection in mice: Comparison of different application methods and correlation of bacterial numbers and histopathology

Munder, Antje; Krusch, Stefan; Tschernig, Thomas; Dorsch, Martina; Lührmann, Anke; Griensven, Martijn van; Tümmler, Burkhard; Weiß, Siegfried; Hedrich, H.-J.

doi:10.1078/0940-2993-00240

Cited by 27 publications

(30 citation statements)

References 20 publications

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“…To verify the correct dilution, an aliquot was serially diluted on LB agar plates. Ten-to 12-week-old female BALB/c mice (Charles River, Sulzfeld, Germany) were inoculated with 30 μl of this bacterial suspension containing 1.0×10 8 CFU or 5×10 6 CFU via view-controlled intratracheal instillation as previously described (Herzog et al 2008;Munder et al 2002). At 1 h or 4 h after the infection, mice were killed by an overdose of normal anesthesia, and then the thorax was opened.…”

Section: Methodsmentioning

confidence: 99%

Bacterial distribution in lung parenchyma early after pulmonary infection with Pseudomonas aeruginosa

et al. 2010

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Nosocomial infections often cause lethal pneumogenic sepsis. Information on early bacteria-host interaction in the lung is limited. In the present study, mice were sacrificed 60 min and 4 h after Pseudomonas aeruginosa (PA) infection to investigate lung morphology by using electron microscopy and light microscopy. After 1 h, bacteria were found in the alveoli partly in contact with surfactant. Alveolar macrophages were seen with up to 10 intracellular bacteria close to protrusions of alveolar epithelial type I cells and the gas/blood barrier. A rare but surprising finding was bacteria and even replicating bacteria in alveolar epithelial type II cells (AEII). No bacteria were seen in capillaries. Neither engulfment of bacteria by neutrophils nor structural damage of the pulmonary barrier was visible. After 4 h, many neutrophils were found within the capillaries, but also in the alveolar space. Thus, we hypothesize that, in early stages of infection, the uptake of PA even by single AEII can influence the course of the disease.

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Section: Methodsmentioning

confidence: 99%

Bacterial distribution in lung parenchyma early after pulmonary infection with Pseudomonas aeruginosa

et al. 2010

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“…Inocula of 6 × 10 5 CFU/30 µl were prepared by dilution with sterile PBS. Anesthetized mice were inoculated via view-controlled intratracheal instillation [17]. …”

Section: Methodsmentioning

confidence: 99%

Head-Out Spirometry Accurately Monitors the Course of <i>Pseudomonas aeruginosa</i> Lung Infection in Mice

Wölbeling

Munder²,

Stanke³

et al. 2010

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Background: Classic infection models in rodents use lethal doses of bacteria as inocula, thus creating models which are rarely comparable to the clinical situation. Moreover, single time-point evaluation requires killing of the animals, necessitating large numbers of animals. Longitudinal parameters such as temperature appear to have a relatively low accuracy. Spirometry might be an accurate method to assess the course of a bacterial lung infection without the necessity to sacrifice the animals. Objectives: We measured lung function in C57BL/6JZtm mice following intratracheal infection with Pseudomonas aeruginosa and compared it to physiological parameters and lung histology. Methods: Head-out spirometry measuring 14 parameters was performed on C57BL6/J mice for eight days following a P. aeruginosa lung infection. Additionally rectal temperature, body weight and condition were assessed together with histological data and bacteriological clearance. Results: Several spirometric parameters were significantly altered for more than 72 h after inoculation, which was four times longer than observed alterations in physiological parameters such as temperature. Volume (amount of air inspired) decreased more than seven-fold within 6 h after inoculation and required 72 h to recover, rendering it the most sensitive spirometric parameter investigated. Spirometric and histological data correlated well. Conclusions: Our findings suggest that non-invasive head-out spirometry is a reliable and highly sensitive method to longitudinally assess the course of bacterial lung infections.

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“…Virulence of the PAO1 sublines was tested by intratracheal instillation of exponentially growing bacteria into the lungs of C3H/HeN mice (28). A dose of 10 6 CFU caused a moderate acute pneumonia by day 2 and led to the death of about half of the animals by day 5 postinfection (Fig.…”

Section: Vol 192 2010mentioning

confidence: 99%

“…Next, we studied the global impact of the sequence diversity among sublines on competitive growth under various in vitro conditions (see Table S2 in the supplemental material) and on virulence in an acute murine airway infection model (28).…”

Section: Vol 192 2010mentioning

confidence: 99%

Genome Diversity ofPseudomonas aeruginosaPAO1 Laboratory Strains

et al. 2010

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Pseudomonas aeruginosa PAO1 is the most commonly used strain for research on this ubiquitous and metabolically versatile opportunistic pathogen. Strain PAO1, a derivative of the original Australian PAO isolate, has been distributed worldwide to laboratories and strain collections. Over decades discordant phenotypes of PAO1 sublines have emerged. Taking the existing PAO1-UW genome sequence (named after the University of Washington, which led the sequencing project) as a blueprint, the genome sequences of reference strains MPAO1 and PAO1-DSM (stored at the German Collection for Microorganisms and Cell Cultures [DSMZ]) were resolved by physical mapping and deep short read sequencing-by-synthesis. MPAO1 has been the source of near-saturation libraries of transposon insertion mutants, and PAO1-DSM is identical in its SpeI-DpnI restriction map with the original isolate. The major genomic differences of MPAO1 and PAO1-DSM in comparison to PAO1-UW are the lack of a large inversion, a duplication of a mobile 12-kb prophage region carrying a distinct integrase and protein phosphatases or kinases, deletions of 3 to 1,006 bp in size, and at least 39 single-nucleotide substitutions, 17 of which affect protein sequences. The PAO1 sublines differed in their ability to cope with nutrient limitation and their virulence in an acute murine airway infection model. Subline PAO1-DSM outnumbered the two other sublines in late stationary growth phase. In conclusion, P. aeruginosa PAO1 shows an ongoing microevolution of genotype and phenotype that jeopardizes the reproducibility of research. High-throughput genome resequencing will resolve more cases and could become a proper quality control for strain collections.The metabolically versatile Pseudomonas aeruginosa is an opportunistic pathogen of plants, animals, and humans and is ubiquitously distributed in soil and aquatic habitats. The common reference strain is P. aeruginosa PAO1, a spontaneous chloramphenicol-resistant mutant of the original PAO strain (earlier called "P. aeruginosa strain 1") that had been isolated in 1954 from a wound in Melbourne, Australia (9, 10). This PAO1 strain from Bruce Holloway's laboratory has become the reference strain for Pseudomonas genetics and functional analyses of the physiology and metabolism of this gammaproteobacterium. A genetic map of its chromosome was generated by exploiting the mechanisms of gene exchange in bacteria, i.e., transduction and conjugation (11). With the advent of pulsedfield gel electrophoresis (PFGE), a physical map of the PAO1 genome was constructed (32) and later merged with the genetic map information (12). By 2000 the PAO1 strain had been completely sequenced (36). Thereafter, the genome annotation has been continually updated and the database content and functionality have been expanded to facilitate accelerated discovery of P. aeruginosa drug targets and vaccine candidates (38). Two near-saturation libraries of transposon insertion mutants have been constructed in P. aeruginosa PAO1 as a global resource for the scient...

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Pulmonary microbial infection in mice: Comparison of different application methods and correlation of bacterial numbers and histopathology

Cited by 27 publications

References 20 publications

Bacterial distribution in lung parenchyma early after pulmonary infection with Pseudomonas aeruginosa

Bacterial distribution in lung parenchyma early after pulmonary infection with Pseudomonas aeruginosa

Head-Out Spirometry Accurately Monitors the Course of <i>Pseudomonas aeruginosa</i> Lung Infection in Mice

Genome Diversity ofPseudomonas aeruginosaPAO1 Laboratory Strains

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